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Overview of strategy for engineering a type IIS restriction endonuclease with a long recognition site. In the first step, the wild-type I-SceI homing endonuclease is mutated into a variant (Sce7) that binds but does not cleave DNA. Second, hybrid enzymes are constructed between Sce7 and the catalytic domain of the type IIS endonuclease FokI, using designed polypeptide linkers.
